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. 2020 Aug 28;31(3):345–361. doi: 10.1038/s41422-020-00397-2

Fig. 1. A non-canonical function of DROSHA in promoting STC1 transcription.

Fig. 1

a Volcano plots displaying DEGs in microarray data comparing shDROSHA with shNC MDA-MB-231 cells. The numbers of significantly variant genes (FC > 1.5, P < 0.05) were shown. Vertical dashed lines indicate cut-off of FC (1.5), whereas the horizontal dashed lines indicate cut-off of P value (0.05). FC, fold change. b Upregulated DEGs in shNC group of microarray data were subjected to GSEA using the gene expression signature that was upregulated in the CD44+/CD24− signature acquired from a public database (GSE7513). c Overlapping stemness gene dataset and downregulated genes in shDROSHA cells from microarray data (FC > 1.5, P < 0.05), common genes were listed. d Relative luciferase activity of pGL3-STC1 in HEK293T cells with forced expression of WT and E1045Q DROSHA. e Western blotting showing STC1 and DROSHA expression in MDA-MB-231 cells with endogenous DROSHA knockdown and forced expression of WT and E1045Q DROSHA. f ChIP was performed to quantify the binding of DROSHA to STC1 in MDA-MB-231 cells with or without 5 μg/mL actinomycin D (ActD). g, h Co-IP assay to detect the interaction between DROSHA and β-Catenin in the nuclear extraction of MDA-MB-231 cells. Histone H3 as nuclear internal control and β-Actin as cytoplasmic internal control. i Relative luciferase activity of pGL3-STC1 in HEK293T cells with knockdown of β-Catenin and forced expression of DROSHA. siβ, siRNA against β-Catenin. j, k Relative luciferase activity of STC1 truncated promoters (j) and mutant promoters (k) in HEK293T cells with overexpression of DROSHA. l The simulated interaction diagram of β-Catenin and DROSHA. m Ability of the indicated Flag-labeled DROSHA derivatives to co-immunoprecipitate His-tagged β-Catenin defined by immunoblotting in HEK293T cells. n Relative luciferase activity of pGL3-STC1 in HEK293T cells with overexpression of the indicated DROSHA derivatives. Data are shown as means ± SD. P values were calculated with two-tailed unpaired Student’s t-test and P < 0.05 is considered statistically significant.